Natural zeolite has been proposed as a dietary additive to enhance broiler performance through modulation of the gastrointestinal environment, detoxification, and improved nutrient utilization. This study evaluated the impact of graduated addition of natural zeolite on growing performance, digestive enzyme activity, serum biochemistry, antioxidant status, hematological parameters, humoral immunity, and detection of NRF2 and TGF-β1 genes in broiler chickens. Three hundred one-day-old unsexed broiler chicks were divided into four food categories at random for 40 days: a control group and three groups supplemented with natural zeolite at 5, 10, and 15 g/kg diet. Zeolite supplementation significantly improved growth performance in a dose-dependent manner, as evidenced by increased live weight and body weight growth, reduced feed consumption, improved conversion of feed rate, and enhanced European Production Efficiency Factor (P<0.05). Digestive enzyme activities (amylase, lipase, and trypsin) were significantly increased (P<0.05). Serum biochemical analysis revealed improved protein metabolism (increased total protein and albumin) and reduced metabolic waste indicators (uric acid and bilirubin) (P<0.05). Antioxidant status was enhanced, with increased total antioxidant capacity and antioxidant enzyme activities (SOD, GSH-Px, and catalase), alongside reduced lipid peroxidation (MDA) (P<0.05). Moreover, the presence of NRF2 and TGF-β1 genes was confirmed in all experimental groups, indicating their potential involvement in antioxidant defense and immune regulation. Hematological parameters showed significant increases in hemoglobin, red blood cell count, and packed cell volume (P<0.05), while white blood cell counts remained stable. Humoral immunity was also improved, as indicated by increased IgM and IgA levels, with IgY showing optimal response at moderate inclusion levels (P<0.05). In conclusion, graded dietary supplementation of natural zeolite improves growth performance, antioxidant capacity, hematological status, and immune function in broiler chickens, potentially through enhancement of intestinal health, nutrient utilization, and modulation of antioxidant and immune-related pathways.

To Cite This Article: Alghamdi RA and Ali AQD, 2026. Effects of natural zeolite on growth performance, antioxidant status, and NRF2 and TGF-β1 gene detection in broilers. Pak Vet J, 46(4): 1008-1017. http://dx.doi.org/10.29261/pakvetj/2026.091